LOCUS       V018968                 7031 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018968
VERSION     V018968
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1324..2901
                     /label="FTO(NM_001363894)"
                     /note="FTO(NM_001363894)"
                     /gene="FTO"
     CDS             2902..2925
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2959..3183
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    3253..3286
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(3744..3760)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        4234..4338
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4339..5199
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      5370..5958
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6246..6267
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6282..6312
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6320..6336
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     6344..6360
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6858..6891
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctgcc
     1321 accatgaagc gcaccccgac tgccgaggaa cgagagcgcg aagctaagaa actgaggctt
     1381 cttgaagagc ttgaagacac ttggctccct tatctgaccc ccaaagatga tgaattctat
     1441 cagcagtggc agctgaaata tcctaaacta attctccgag aagccagcag tgtatctgag
     1501 gagctccata aagaggttca agaagccttt ctcacactgc acaagcatgg ctgcttattt
     1561 cgggacctgg ttaggatcca aggcaaagat ctgctcactc cggtatctcg catcctcatt
     1621 ggtaatccag gctgcaccta caagtacctg aacaccaggc tctttacggt cccctggcca
     1681 gtgaaagggt ctaatataaa acacaccgag gctgaaatag ccgctgcttg tgagaccttc
     1741 ctcaagctca atgactacct gcagatagaa accatccagg ctttggaaga acttgctgcc
     1801 aaagagaagg ctaatgagga tgctgtgcca ttgtgtatgt ctgcagattt ccccagggtt
     1861 gggatgggtt catcctacaa cggacaagat gaagtggaca ttaagagcag agcagcatac
     1921 aacgtaactt tgctgaattt catggatcct cagaaaatgc catacctgaa agaggaacct
     1981 tattttggca tggggaaaat ggcagtgagc tggcatcatg atgaaaatct ggtggacagg
     2041 tcagcggtgg cagtgtacag ttatagctgt gaaggccctg aagaggaaag tgaggatgac
     2101 tctcatctcg aaggcaggga tcctgatatt tggcatgttg gttttaagat ctcatgggac
     2161 atagagacac ctggtttggc gatacccctt caccaaggag actgctattt catgcttgat
     2221 gatctcaatg ccacccacca acactgtgtt ttggccggtt cacaacctcg gtttagttcc
     2281 acccaccgag tggcagagtg ctcaacagga accttggatt atattttaca acgctgtcag
     2341 ttggctctgc agaatgtctg tgacgatgtg gacaatgatg atgtctcttt gaaatccttt
     2401 gagcctgcag ttttgaaaca aggagaagaa attcataatg aggtcgagtt tgagtggctg
     2461 aggcagtttt ggtttcaagg caatcgatac agaaagtgca ctgactggtg gtgtcaaccc
     2521 atggctcaac tggaagcact gtggaagaag atggagggtg tgataagtgt ggagttatac
     2581 aaagagctag gacttatgtc tcccaggttg ataaggcaca aggcaacaaa tgctgtgctt
     2641 catgaagtta aaagagaggg gctccccgtg gaacaaagga atgaaatctt gactgccatc
     2701 cttgcctcgc tcactgcacg ccagaacctg aggagagaat ggcatgccag gtgccagtca
     2761 cgaattgccc gaacattacc tgctgatcag aagccagaat gtcggccata ctgggaaaag
     2821 gatgatgctt cgatgcctct gccgtttgac ctcacagaca tcgtttcaga actcagaggt
     2881 cagcttctgg aagcaaaacc cgattacaag gatgacgacg ataagtgacc atggtttaaa
     2941 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3001 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3061 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3121 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3181 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3241 gcggcgcatt aaataacttc gtataatgta tgctatacga agttatatcg tatgcatgct
     3301 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     3361 ccacagaatt ccttgaaata catccacagg tacacctcca attgactcaa atgatgtcaa
     3421 ttagtctatc agaagcttct aaagccatga catcattttc tggaattttc caagctgttt
     3481 aaaggcacag tcaacttagt gtatgtaaac ttctgaccca ctggaattgt gatacagtga
     3541 attataagtg aaataatctg tctgtaaaca attgttggaa aaatgacttg tgtcatgcac
     3601 aaagtagatg tcctaactga cttgccaaaa ctattgtttg ttaacaagaa atttgtggag
     3661 tagttgaaaa acgagtttta atgactccaa cttaagtgta tgtaaacttc cgacttcaac
     3721 tgtagggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3781 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3841 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3901 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3961 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4021 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4081 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4141 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4201 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4321 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4381 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4441 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4501 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4561 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4621 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4681 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4741 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4801 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4861 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4921 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4981 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5041 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5101 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5161 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6361 catgattacg ccaagcttgc atgcctgcag gtcgactcta gacctataca gttgaagtcg
     6421 gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta ctccacaaat
     6481 ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt gtgcatgaca
     6541 caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat tcactgtatc
     6601 acaattccag tgggtcagaa gtttacatac actaagttga ctgtgccttt aaacagcttg
     6661 gaaaattcca gaaaatgatg tcatggcttt agaagcttct gatagactaa ttgacatcat
     6721 ttgagtcaat tggaggtgta cctgtggatg tatttcaagg ggatccagac atgataagat
     6781 acattgatga gtttggacaa accacaacta gaatgcagcg gccgctagct cgagatctta
     6841 attaaactag tatcgatata acttcgtata atgtatgcta tacgaagtta tatgtacggg
     6901 ccagatatac gcgttgacat tgattattga ctaggctttt gcaaaaagct ttgcaaagat
     6961 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
     7021 gagtgggaat t
//
